validation guide summary - biopharm.saint-gobain.com · november 18, 2013 telephone (989) 435-9533...

FLS-5201VS 3910 Terry-Diane Street, Beaverton, MI 48612 November 18, 2013 Telephone (989) 435-9533 Fax (989) 435-2355

Validation Guide Summary

Tygon® 3355L Revision: November 18, 2013


Tygon® 3355L Validation Guide Summary

Table of Contents Page 1. Summary _____________________________________________________________________________________ 3

2. Typical Physical Properties _______________________________________________________________________ 3

3. Regulatory and Information Overview ______________________________________________________________ 4

a. Country of Origin _______________________________________________________________________ 4

b. Animal Derivative Content Free ____________________________________________________________ 4

c. US Food and Drug Administration (FDA) _____________________________________________________ 4

d. Metals with Safety Concern (EMEA/CHMP/SWP/4446/2000) ____________________________________ 4

e. Coalition of Northeastern Governors (CONEG) ________________________________________________ 5

f. Restriction of Hazardous Substances (RoHS) __________________________________________________ 5

g. European Directive 2003/22/EC ____________________________________________________________ 5

h. California Proposition 65 _________________________________________________________________ 5

i. Phthalates _____________________________________________________________________________ 5

j. Perfluorinated Compounds _______________________________________________________________ 6

k. Ozone Depleting Chemicals _______________________________________________________________ 6

l. PBA __________________________________________________________________________________ 6

m. Melanine ______________________________________________________________________________ 6

n. Latex, Gluten and Allergens _______________________________________________________________ 6

o. REACH ________________________________________________________________________________ 6

p. Shelf Life ______________________________________________________________________________ 7

4. Biocompatibility, Physiochemical and Extractable Testing ______________________________________________ 8

a. Summary ______________________________________________________________________________ 8

b. Hemolysis: Direct Contact ________________________________________________________________ 9

c. Hemolysis: Indirect Contact _______________________________________________________________ 9

d. Irritation and Skin Sensitization – Kligman Maximization _______________________________________ 10

e. Systemic Toxicity – Rabbit Pyrogen (Material Mediated) _______________________________________ 10

f. Systemic Toxicity – Systemic Injection ______________________________________________________ 11

g. Biological Reactivity Tests, In Vitro – Direct Contact ___________________________________________ 11

h. Biological Reactivity Tests, In Vitro – MEM Elution ____________________________________________ 11

i. Biological Reactivity Tests, In Vitro – Agar Diffusion ___________________________________________ 12

j. Biological Reactivity Tests, In Vivo _________________________________________________________ 12

k. Pyrogen Test __________________________________________________________________________ 13

l. Elastomeric Closures for Injection _________________________________________________________ 13

m. Total Organic Carbon Analysis ____________________________________________________________ 14

n. Physiochemical Tests – Plastics ___________________________________________________________ 14

o. Total Extractables in Rubber Articles Intended for Repeated Use ________________________________ 15

p. Silicone Elastomer for Closures and Tubing __________________________________________________ 16

q. Metals Analysis buy ICP-MS: Analysis for Arsenic, Cadmium, Lead and Mercury ____________________ 16


Validation Guide Summary: Tygon® 3355L 1.0 Summary

Tygon® 3355L is an ultra-pure biopharmaceutical-grade platinum-cured silicone tubing. It provides unsurpassed inner surface

smoothness and is ultra-flexible. Tygon® 3355L lasts up to five times longer than other platinum silicone tubings. It has low

extractables to prevent product contamination. It imparts no tastes or odors to fluids. Tygon® 3355L is biocompatible and has

excellent fluid flow characteristics.

Tygon 3355L meets all relevant USP Class VI requirements.

2.0 Typical Physical Properties

Property ASTM Standard Value or Rating

Durometer Hardness , Shore A, 15 Sec D2240-97 55

Color — Translucent

Tensile Strength , psi (MPa) D412-97 900 (6.2)

Ultimate Elongation, % D412-97 400

Tear Resistance, lb-f/inch (kN/m) D624-91, Die B 135 (23.6)

Specific Gravity D792-91 1.12

Water Absorption, % , 24 hrs. @ 23°C D570-95 0.16

Compression Set

Constant Deflection, % @158°F (70°C) for

22hrs

D395-89

Method B 5 5

Brittleness by Impact, Temp., °F (°C) D746-95 <-112°F (<-80°C)

Maximum Recommended —

Operating Temp.,°F (°C) — 400 (204)

Dielectric Strength, v/mil (kV/mm) D149-95 462 (18.2)

Tensile Modulus,

@ 100% Elongation, psi (MPa)

@ 200% Elongation, psi (MPa)

D412-97

375 (2.6)

500 (3.4)

Tensile Set, % D412-97 6


3.0 Regulatory Information Overview - Effective Date: June 7, 2013

Thank you for your interest in Saint-Gobain’s Tygon® platinum-cured silicone tubing, hose, manifolds, TAs, profiles, and

molded products and services that we provide.

Please be advised that Tygon® Silicone Products are manufactured from 100% synthetic virgin silicone. No plant or animal

products or by-products are used in the manufacture of nor intentionally added.

Saint-Gobain Performance Plastics Beaverton’s (SGPPL-BV) Tygon® Silicone Products are manufactured in an ISO 14644

Class 7 clean room facility from USP 88 Class VI and FDA 21CFR approved raw materials in accordance with cGMP principles.

3a. Country of Origin

SGPPL-BV’s Tygon® Silicone Products are manufactured in the U.S.A

3b. Animal Derivative Content Free - Tallow/BSE/TSE

Please be advised that we have found no instance of the use of bovine or other animal derived materials used in the

manufacture of any Tygon® Silicone Products as defined in specification EMEA 410.01 Rev.3.

3c. US Food and Drug Administration (FDA):

All materials used to produce Tygon® Silicone Products meet the FDA requirements outlined in the Code of Federal

Regulations 21 CFR 177.2600(a) and (b). This conformance includes all ingredients used in the product formulation.

These ingredients are compliant to their specific regulations and 21 CFR 177.2600(c). This product can be used for

food contact applications with food types I, II, IV-B, VI, VII-B, and VIII of Table 1 and under conditions of use C through

H of Table 2 in 21 CFR 176.170(c).

3d. Metals with Safety Concern (EMEA/CHMP/SWP/4446/2000)

Tygon® Silicone Product formulations were tested by ICP-MS to a sensitivity of 1 ppm. The quantity of Metals with

Safety Concerns present in Tygon® Silicone Products is deemed below the Parenteral Exposure limits given in

EMEA/CHMP/SWP/4446/2000 which establishes:

Classification Metals Parenteral Exposure Limit

Metals of significant safety concern

Class 1A Pt, Pd 1 ppm

Class 1B* Ir, Rh, Ru, Os 1 ppm

Class 1C Mo, Ni, Cr, V 2.5 ppm

Metals with low safety concern Class 2 Cu, Mn 25 ppm

Metals with minimal safety concern Class 3 Fe, Zn 130 ppm


*While Class 1B is a subclass limit of 1 ppm, based on the formulation, supplier information, the metals in this subclass testing below the sensitivity of the

test, and the fact that these metals are not, and will not be used in the manufacture of, or intentionally added to Tygon ® Silicone Products, we would not

expect the total of these materials to be present in quantities greater than 1 ppm.

3e. Coalition of Northeastern Governors (CONEG)

Cadmium, chromium, lead, and mercury are not used in the manufacture or formulation of Tygon® Silicone Products. In

addition, this product meets the CONEG limitation of 100 ppm for the total incidental content of cadmium, chromium, lead,

and mercury.

3f. Restriction of Hazardous Substances (RoHS)

Saint-Gobain Performance Plastics, in Compliance with the European Union restriction of the use of certain Hazardous

Substances (RoHS) directives 2011/65/EU, 2002/96/EC, 2003/11/EC, hereby certifies that the following materials or

substances are not, and will not be used in the manufacture of, or intentionally added to Tygon ® Silicone Products: Lead

(Pb), Mercury (Hg), Cadmium (Cd), Hexavalent Chromium (CrVI), and two Bromine containing Flame Retardants – PBB

(Polybrominated Biphenyls) and PBDE (Polybrominated Diphenyl Ethers) that are in concentrations greater than 0.1% by

mass (1,000 ppm). Cadmium (CrVI) concentration limit to be not greater than 0.01% (100 ppm). In addition, we would not

expect the total of these materials to exceed 100 ppm (0.0 1% by mass).

3g. European Directive 2003/11/EC

Tygon® Silicone Products conform to the European Directive 2003/11/EC of 6 February 2003 amending 76/769/EEC of 27

July 1976. We do not use Polychlorinated biphenyls (PCB), polychlorinated terphenyls (PCT), Vinyl Chloride, Pentabromo-

diphenyl ethers (pentaBDE), Octabromo-diphenyl ethers (OctaBDE), or Decabromo-diphenyl ethers (decaBDE) in the

manufacture or formulation of this product.

3h. California’s Safe Drinking Water and Toxic Enforcement Act of 1986 (Proposition 65)

Based on available information Tygon® Silicone Products do not contain any components or chemicals currently known to the

State of California to cause cancer, birth defects or reproductive harm at levels which would be subject to Proposition 65.

3i. Phthalates

Phthalates are not and will not be used in the manufacture of nor intentionally added to the process or raw materials of

Tygon® Silicone Products, including, but not limited to:

Bis(2-ethylhexyl)phthalate (DEHP), Bis(2-methoxyethyl) phthalate (DMEP), Dibutyl phthalate (DBP), benzxyl butyl phthalate

(BBP), Diisononyl Phthalate (DINP), Diisodecyl phthalate (DIDP), Didecyl Phthalate (DDP), Di-n-hexyl phthalate (DnHP), Di-n-

pentyl phthalate (DnPP), Diisopentyl phthalate (DIPP), Diisobutyl phthalate (DIBP), n-Pentyl-isopentyl phthalate (nPIP),

Di-n-octyl phthalate (DnOP).


3j. Perfluorinated Compounds

Perfluorooctanoic Acid (PFOA) or Perfluorooctane sulfantes (PFOS), is not used in the formulation or manufacture of

Tygon® Silicone Products.

3k. Ozone Depleting Chemicals (ODCs)

We do not use Class I or Class II ODCs listed in the Resolution 2037/2000/EC in the manufacture or formulation of

Tygon® Silicone Products.

3l. BPA

Tygon® Silicone Products do not have Bisphenol A introduced in the process or the raw materials.

3m. Melamine

Melamine is not and will not be used in the manufacture of nor intentionally added to the process for any of our Tygon®

Silicone Products. 3910 Terry-Diane Street Beaverton, MI 48612 Telephone (989) 435-9533 Fax (989) 435-2355

3n. Latex, Gluten, and Allergens

Please be advised that all Tygon® Silicone Products neither contain latex or any form of latex material, nor does the

manufacturing process and equipment utilize or contact latex materials. In addition, the materials used in Tygon®

Silicone Products do not contain other allergens - as defined by FDA as Milk, Eggs, Fish, Crustaceans, Soy, Wheat or

other Gluten containing sources, Peanuts and Tree Nuts – neither in the manufacture nor the formulation. Furthermore,

there is no instance where these allergens come in contact with the finished product during any step of the

manufacturing process.

3o. REACH

Saint-Gobain PPL has met the requirements set forth by REACH legislation as it relates to pre-registration of certain

substances by December 1, 2008. We will continue to monitor regulatory activity in this area to be certain that we stay

abreast of all requirements and comply as needed.

Saint-Gobain Performance Plastics complies with provisions of REACH where applicable to our products. Tygon® Silicone

Products qualify as articles as defined by REACH legislation. Furthermore, no substances are intentionally released as a

result of using our products in the intended manner.

Substance of Very High Concern (SVHC) as established by the European Union/ECHA Candidate list posted December 19,

2012 are not intentionally added or formulated in Tygon® Silicone Products or their base resins. We will continue to


monitor regulatory activity in this area to be certain that we stay abreast of all requirements and comply as needed. Updated

SVHC compliance statements can be obtained from Tygon® Silicone Product suppliers, as necessary.

3p. Shelf Life

The shelf life of Tygon® Silicone Products are 5 years from date of manufacture, provided the product is packaged in the

original unopened packaging and is stored under ambient temperature and humidity conditions.

Use of this Regulatory Information Overview

The information in this Regulatory Information Overview is provided for the purpose of communicating regulatory

compliance of Tygon® silicone tubing and hose products as shipped from a Saint-Gobain location. Ultimately, customers

must make their own determination that use of this product is safe, lawful, and technically suitable for their intended

applications.

Saint-Gobain has not run any analysis for concentration levels for the regulatory compliances listed above but has relied on

raw material suppliers for this information. Based upon supplier information, we would expect that any trace amounts of the

chemicals contained in the above compliance statements to be at concentrations levels that would be far below the

allowable concentration limits.


4.0 Tygon® 3355L Biocompatibility, Physiochemical & Extractable Testing

4a. Summary The following is a summary of the validation testing that has been performed on Tygon® 3355L Tubing. Complete test reports can be found in the Tygon® 3355L Validation Guide.

Test Standard Result

Hemolysis – Direct Contact ISO 10993-4 Passed

Hemolysis – Indirect Contact ISO 10993-4 Passed

Irritation and Skin Sensitization: Kligman Maximization Test ISO 10993-10 Passed

Systemic Toxicity - Rabbit Pyrogen (Material Mediated) ISO 10993-11 Passed

Systemic Toxicity – Systemic Injection Test ISO 10993-11 Passed

Biological Reactivity Tests, In Vitro: Direct Contact USP <87> Passed

Biological Reactivity Tests, In Vitro: MEM Elution USP <87> Passed

Biological Reactivity Tests, In Vitro: Agar Diffusion USP <87> Passed

Class VI Plastics - Biologic Reactivity Tests, In Vivo USP <88> Passed

Pryogen Test USP <151> Passed

Elastomeric Closures for Injection, see Summary, 4l. USP <381>

Total Organic Carbon USP <643> 19.8 mg/l

Physicochemical Tests for Plastics USP <661> Passed

Rubber Closures for Containers EP 3.2.9 Passed

Total Extractables in Rubber Articles Intended for Repeated Use 21 CFR Part 117.2600 Passed

Silicone Elastomer for Closure and Tubing EP 3.1.9 Passed

Metal Analysis via ICP-MS, see Summary, 4q


4b ISO 10993-4 Hemolysis: Direct Contact

The Hemolysis test assesses the potential for direct contact of a given sample with blood to cause the rupture of

erythrocytes (red blood cells).

Test: A sample of Tygon® 3355L was tested by Toxikon Corporation in accordance with ISO 10993-4, 2002, Biological

Evaluation of Medical Devices – Part 4: Selection of Tests for Interactions with Blood, as amended 2006.

The test article was added to a vial containing USP 0.9% Sodium Chloride for Injection (NaCl). The test article extract, the

positive control (USP Sterile Water for Injection) and negative control (NaCl) were then incubated in a waterbath at 37oC

for 30 minutes. Rabbit blood was then added to each vial and the resulting mixture was incubated in a water bath at 37oC

for a further 60 minutes. The incubated vials were then centrifuged and the supernatant was drawn off. The absorbance

of the supernatant samples was then determined via spectrophotometer at 545nm to determine the corresponding

hemoglobin concentration.

Results: The percent hemolysis resulting from direct contact of the product with rabbit blood was 0.0% above the

negative control. Per ISO 10993-4, a test article is considered non-hemolytic if its percent hemolysis is <5.0% above the

negative control. The test article was therefore deemed non-hemolytic.

4c. ISO 10993-4 Hemolysis: Indirect Contact

The Hemolysis test assesses the potential for indirect contact of a given sample with blood to cause the rupture of

erythrocytes (red blood cells).


Evaluation of Medical Devices – Part 4: Selection of Tests for Interactions with Blood.

Test samples and controls were prepared by adding a sample of the test article to vials of Phosphate Buffered Saline (PBS).

Sterile Water for Injection was used as the positive control and PBS was used as the negative control. Vials of test article

extract, positive control and negative control samples were incubated in a water bath at 37oC for 30 minutes. After the

incubation period, 0.2ml of rabbit blood was added to each vial, and the vials were then incubated for a further 60

minutes at 37oC. The samples were then centrifuged and the supernatant was drawn off. The absorbance of the samples

was measured at 545nm to determine the corresponding hemoglobin concentration.

Results: The percent hemolysis resulting from indirect contact of the product with rabbit blood was 0.2% above the

negative control. Per ISO 10993-4, a test article is considered non-hemolytic if its percent hemolysis is <5.0% above the

negative control. The test article was therefore deemed non-hemolytic.


4d. ISO 10993-10 Tests for Irritation and Skin Sensitization: Kligman Maximization Test

The tests for irritation and skin sensitization are performed to determine the allergenic potential of a test article on

contact with skin.


Evaluation of Medical Devices – Part 10: Tests for Irritation and Skin Sensitization, as amended 2006.

Samples of the test article were extracted in USP 0.9% Sodium Chloride (NaCl) or Cottonseed Oil at 70oC for 24 hours. 35

Hartley guinea pigs (20 experimental, 10 negative control, 5 positive control) were used for these evaluations. On Day 0

(Induction Phase) samples of the test article extracts and positive and negative controls were injected intradermally into

the test animals. On Day 7 (Topical Application Phase) the test article extracts and control articles were applied to the test

sites and left in place for 48 hours. On Day 23 (Challenge Phase) the test article extracts and control articles were applied

to the test sites and left in place for 24 hours. The test and control animals were examined and scored for erythema and

edema according to the Magnusson and Kligman Scale at 24, 48 and 72 hours post Challenge Phase.

Results: The skin sites exposed to the test article extracts and the negative control showed no evidence of erythema or

edema. The sites exposed to the positive control demonstrated erythema and edema as expected. The test article extracts

therefore produced no reaction (0% sensitization) in the test animals, corresponding to a Kligman Grade I reaction and

indicating that the test article can be classified as having a weak allergenic potential.

4e. ISO 10993-11 Tests for Systemic Toxicity – Rabbit Pyrogen Test (Material Mediated)

The rabbit pyrogen test is performed to qualitatively determine whether a given test article contains pyrogens. Pyrogens

can provoke a significant febrile reaction in a human patient who receives a parenteral drug product that has come into

contact with a contaminated test article.

Test: A sample of Tygon® 3355L was tested by Toxikon Corporation in accordance with ISO 10993-11:2006 Biological

Evaluation of Medical Devices Part 11, Tests for Systemic Toxicity.

The test article was immersed in USP 0.9% Sodium Chloride for Injection (NaCl) at 70oC for 24hrs. The resulting extract

was administered to test subjects (rabbits) via IV injection at a dose of 10 mL per kg of body mass. A control rabbit was

similarly injected with the control sample (NaCl). The body temperatures of the animals were measured 30 minutes prior

to injection and again every 30 minutes between the 1 hour and 3 hours marks post-injection.

Results: The three test rabbits showed changes of 0.0oC, 0.2

oC and 0.4

oC in body temperature post-injection. The control

rabbit showed a 0.0oC change in body temperature post-injection. A test article is considered pyrogen-free provided that

none of the test subjects display an increase in body temperature of more than 0.5oC post-injection, therefore the Tygon®

3355L test articles were deemed pyrogen-free.


4f. ISO 10993-11 Tests for Systemic Toxicity – Systemic Injection

The systemic injection test is performed to assess the potential for a given test article extract to cause toxic effects as a

result of a single dose systemic injection. The test is performed using mice.

Test: A sample of Tygon® 3355L was tested by Toxikon Corporation in accordance with ISO 10993-11:1993 Biological

Evaluation of Medical Devices Part 11, Tests for Systemic Toxicity.

Test articles were immersed in USP 0.9% Sodium Chloride (NaCl), Cottonseed Oil (CSO), 1 in 20 Ethanol in NaCl (EtOH) or

Polyethylene Glycol 400 (PEG) at 121oC for 1 hour. The test article extracts and corresponding controls (samples of each

extractant that had not been exposed to the test article) were injected intravenously (NaCl and EtOH) or intraperitoneally

into Albino Swiss Mice and the animals were observed for 72 hours for signs of biological reactivity.

Results: The animals treated with the test article extracts did exhibit a significantly greater biological reaction than the

negative controls toxicity during the observation period. The test article is therefore considered non-toxic per the criteria

of the ISO 10993-11 Systemic Injection test.

4g. USP <87> Biological Reactivity Tests, In Vitro – Direct Contact

Cytotoxicity testing assesses the potential of a given material to have a toxic effect on living cells.

Test: Samples of Tygon® 3355L were tested by Toxikon Corporation in accordance with USP 26, NF 21, 2003; <87>

Biological Reactivity Tests, In Vitro.

The test article sample, negative control (Negative Control Plastic) sample and positive control (Natural Rubber) sample

were placed directly onto mammalian monolayers of L929 mouse fibroblast cells. The cultures were incubated at 37oC in

5% CO2 for 48hrs. Cultures were monitored for cellular degeneration and malformation and rated on a scale of 0 (No

Biological Reactivity) to 4 (Severe Biological Reactivity).

Results: The test article samples scored a Grade 0 for Biological Reactivity after 48hrs. The negative controls scored a

Grade 0. The positive controls scored a Grade 3 at the 48hr mark. Samples are deemed to meet the test requirements if

they exhibit a Biological Reactivity of no more than Grade 2 (Mild Reactivity). The test articles are therefore considered

non-cytotoxic.

4h. USP <87> Biological Reactivity Tests, In Vitro - MEM Elution Test





Test samples were immersed in Serum-Supplemented Minimum Essential Medium at 37oC for 24hrs. Positive control

(Natural Rubber) and negative control (Negative Control Plastic) samples were also extracted as above. Duplicates of all

three extracts were incubated with L929 mouse fibroblast cells at 37oC for 48hrs. Cultures were monitored for cellular

degeneration and malformation and rated on a scale of 0 (No Biological Reactivity) to 4 (Severe Biological Reactivity).

Results: The test article samples and the negative controls scored a Grade 0 for Biological Reactivity after 48hrs. The

positive controls scored a Grade 4 at the 48hr mark. Samples are deemed to meet the test requirements if they exhibit a

Biological Reactivity of no more than Grade 2 (Mild Reactivity). The test articles are therefore considered non-cytotoxic.

4i. USP <87> Biological Reactivity Tests, In Vitro – Agar Diffusion Test




Duplicate test article, negative control (Negative Control Plastic) and positive control (Buna-N-Rubber) samples were

placed in separate wells containing solidified agarose stained with vital dye (neutral red) overlaying a L929 mouse

fibroblast monolayer. The culture plates were then incubated at 37oC in 5% CO2 for 24hrs. After the incubation period

cultures were examined macroscopically and microscopically for cell decolorization and potential cell lysis and rated on a

scale of 0 (No Biological Reactivity) to 4 (Severe Biological Reactivity).

Results: The test article samples and the negative controls scored a Grade 0 for Biological Reactivity after 48hrs. The

positive controls scored a Grade 3 at the 48hr mark. Samples are deemed to meet the test requirements if they exhibit a

Biological Reactivity of no more than Grade 2 (Mild Reactivity). The test articles are therefore considered non-cytotoxic.

4j. USP <88> Biological Reactivity Tests, In Vivo

The USP Class VI Plastics Test assesses the potential toxicity of a given test article by introducing a sample into live animals

systemically, intracutaneously and through implantation. Test animals are then monitored for signs of irritation and/or

toxicity.

Test: Samples of Tygon® 3355L were tested by Toxikon Corporation in accordance with USP 29, NF 24, 2006, <88>

Biological Reactivity Tests, In Vivo.

Test articles were immersed in USP 0.9% Sodium Chloride (NaCl), Cottonseed Oil (CSO), 1 in 20 Ethanol in NaCl (EtOH) or

Polyethylene Glycol 400 (PEG) at 70oC for 24 hours. The test article extracts and corresponding controls (samples of each

extractant that had not been exposed to the test article) were injected systemically into mice and intracutaneously into

rabbits and the animals were observed for 72 hours for signs of skin reactivity or toxicity. In addition, the test article was

implanted into the paravertebral muscles of rabbits, which were then observed for 7 days for macroscopic signs of

hemorrhage, necrosis, discoloration, encapsulation and/or infection.


Results: None of the animals injected systemically with test article extracts or controls exhibited any signs of toxicity.

Similarly, none of the animals injected intracutaneously with test article extracts or controls exhibited any signs of

erythema, edema or clinical toxicity. Further, none of the implanted animals exhibited any signs of toxicity at the

implantation sites relative to the control sites. The test article therefore met the requirements of the USP Class VI Test for

Biocompatibility.

4k. USP <151> Pyrogen Test

The rabbit pyrogen test is performed to qualitatively determine whether a given test article contains pyrogens. Pyrogens

can provoke a significant febrile reaction in a human patient who receives a parenteral drug product that has come into

contact with a contaminated test article.

Test: A sample of Tygon® 3355L was tested by Toxikon Corporation in accordance with USP 26, NF 21, 2003; <151>

Pyrogen Test.

The test article was immersed in USP 0.9% Sodium Chloride for Injection (NaCl) at 70oC for 24hrs. The resulting extract

was administered to test subjects (rabbits) via IV injection at a dose of 10 mL per kg of body mass. A control rabbit was

similarly injected with the control sample (NaCl). The body temperatures of the animals were measured 30 minutes prior

to injection and again every 30 minutes between the 1 hour and 3 hours marks post-injection.

Results: The test rabbits showed changes in body temperature post-injection of 0.1oC, 0.2

oC and 0.0

oC. There was no

change in body temperature in the control rabbit post-injection. A test article is considered pyrogen-free provided that

none of the test subjects display an increase in body temperature of more than 0.5oC post-injection, therefore the test

articles were deemed pyrogen-free.

4l. USP <381> Elastomeric Closures for Injection

Testing of elastomeric closures for use with containers for injectables is performed to assess the suitability of the test

article for use in contact with drug products for parenteral administration in humans.


Elastomeric Closures for Injections.

The test article extracts were prepared by immersing duplicate test article samples in purified water and autoclaving at

121oC for 30 minutes, then rinsing twice with purified water. One of the duplicate test articles was subsequently

immersed in 200 ml of fresh purified water and autoclaved at 121oC for 2 hours. The other test article sample was

extracted in 200 ml of Isopropyl Alcohol (IPA) by reflux apparatus for 2 hours. The test extracts were then tested for

Turbidity, Reducing Agents, Heavy Metals, pH Change and Total Extractables per USP <381>.

Results: The test results are summarized in the table below.


Parameter Results

Water IPA

Turbidity 1.25 NTU 0.05 NTU

Total Extractables 0 mg 48 mg

Reducing Agents 0.02 ml Not applicable

Heavy Metals ≤ 1.0 ppm Not applicable

pH Change 0 pH units Not applicable

4m. USP <643> Total Organic Carbon Analysis

The test for Total Organic Carbon provides an indirect measure of the total organic matter present in a given sample.

Test: A sample of Tygon® 3355L was tested by Toxikon Corporation in accordance with USP 31, NF 26, 2008; <643> Total

Organic Carbon.

The test article was immersed in Purified Water at 70oC for 24hrs. The Total Organic Carbon in the test article extract was

determined by converting TOC to carbon dioxide through acidification and chemical wet oxidation with sodium persulfate.

The CO2 released by the extract was measured using an infrared detector.

Results: The results of the test are summarized in the table below.

Sample Results Detection Limit

(mg/l) (mg/l) (mg/cm)

Tygon® 3355L 19.8 0.01 0.1

4n. USP <661> Physicochemical Tests for Plastics

Physicochemical testing is performed to assess the suitability of the test article for use in contact with drug products for

parenteral administration in humans.


Containers, Physicochemical Tests – Plastics.


The test article was immersed in USP Purified Water for 70oC for 24hrs. The extract was then tested for Non-Volatile

Residue, Residue on Ignition, Heavy Metals as Lead and Buffering Capacity per USP <661>.


Assay Assay Results Limits Based on Area

Non-Volatile Residue 0.0 mg ≤ 15 mg

Residue on Ignition Not applicable ≤ 5 mg

Heavy Metals < 1 ppm ≤ 1 ppm

Buffering Capacity 0.050 ml ≤ 10 ml

The test article sample met the criteria established per USP <661> for all of the tests performed, as shown above.

4o. 21 CFR Part 117.2600 Total Extractables in Rubber Articles Intended for Repeated Use

The Total Extractables Test is intended to determine the total solids per unit surface area that can be extracted from a

given test article using purified water and hexane.

Test: A sample of Tygon® 3355L was tested by Toxikon Corporation in accordance with 21 CFR Part 177.2600, 2006:

Rubber Articles Intended for Repeated Use.

The test articles were immersed in either purified water or hexane for 7hrs (the Reflux Time). The test articles were then

removed and extracted with fresh samples of purified water and n-hexane for a further 2hrs. The resulting extracts were

then evaporated to dryness and the residue per unit surface area was determined for each sample.


Reflux Time Purified Water

Results Evaluation Criteria Meets Criteria?

First 7 hours 0.00 mg/in2 ≤ 20 mg/in

2 Yes

Second 2 Hours 0.00 mg/in2 ≤ 1 mg/in

2 No

Reflux Time N-Hexane

Results Evaluation Criteria Meets Criteria?

First 7 hours 19.3 mg/in2 ≤ 175 mg/in

2 Yes

Second 2 Hours 3.50 mg/in2 ≤ 4 mg/in

2 No


As shown in the table above, the test article samples met the criteria established per 21 CFR Part 177.2600: Rubber

Articles Intended for Repeated Use.

4p. EP 3.1.9 Silicone Elastomer for Closures and Tubing

The European Pharmacopoeia Monograph Testing Section 3.1.9 assesses the physical and physicochemical characteristics

of silicone elastomer closures and tubing.

Test: Samples of Tygon® 3355L were tested by Toxikon Corporation in accordance with EP 4.0, 2002, Section 3.1.9.

The test articles were examined according to the test procedures specified in EP 3.1.9. Test results were assessed relative

to the specified EP limit for each characteristic.


Parameter Results Acceptance Limits Meets Criteria?

Appearance Clear, Turbidity < RSI Clear, Turbidity < RSI Yes

Acidity or Alkalinity Pass

Pass

≤ 2.5 ml of 0.01 M NaOH

≤ 1.0 ml of 0.01 M HCl Yes

Relative Density 1.13 g/ml Between 1.05 and 1.25 g/ml Yes

Reducing Substances 0.0 ml Difference between titration

volumes ≤ 1.0 ml Yes

Substances Soluble in

Hexane 14.1 mg ≤ 15.0 mg (or 3 %) Yes

Volatile Matter 0.158 % ≤ 2.0 % for platinum-cured Yes

Mineral Oils No fluorescence Less fluorescence than 1 ppm

Quinine Sulfate standard Yes

Phenylated Compounds < 0.1 mAU Absorbance ≤ 0.4 mAU from 250

nm and 340 nm Yes

Platinum No color Less color than standard Yes

The test article samples met the criteria established per EP 3.1.9 for all of the tests performed, as shown above.

4q. Metals Analysis by ICP-MS: Analysis for Arsenic, Cadmium, Lead and Mercury

Elemental impurities can be highly toxic to humans even at very low concentrations. An analysis of the concentration of

individual elements in a product intended for use in biopharmaceutical applications must therefore be performed.


Test: A sample of Tygon® 3355L was tested by Toxikon Corporation for the presence of heavy metals via elemental analysis

by ICP (inductively-coupled plasma), in accordance with EPA Methods 6010A and 7471.

The test article was digested with nitric acid in a microwave and then analyzed via Inductively Coupled Plasma – Mass

Spectroscopy. A separate test article sample was digested in aqua regia, oxidized with potassium permanganate and

followed by reaction by hydroxylamine. The resulting sample was analyzed for mercury via Cold Vapor Atomic Absorption

(CVAA) Spectroscopy.

Results: The results are provided in the table below.

Element

Results

g/g

Detection Limit

g/g

Arsenic ND 5.00

Cadmium ND 0.50

Lead ND 2.50

Mercury ND 0.083

validation guide summary - biopharm.saint-gobain.com · november 18, 2013 telephone (989) 435-9533...

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